Medium data

Search for medium no. = [1030].

Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1030 DISSULFURIMICROBIUM HYDROTHERMALE MEDIUM

NH₄Cl	`0.33`	g
KCl	`0.33`	g
MgCl₂·6H₂O	`0.33`	g
CaCl₂·2H₂O	`0.33`	g
KH₂PO₄	`0.33`	g
Trace elements solution (see below)	`10.0`	ml
Sulfur (powder)	`5.0`	g
Ferrihydrite suspension (see below)	`200.0`	ml
Distilled water	`800.0`	ml

Mix components, except ferrihydrite suspension and sulfur, bring to a boil and cool down under a CO₂ (100%) gas stream. While gassing, add the following components to the medium:

Trace vitamin (see Medium No. 197)	`10.0`	ml
NaHCO₃	`2.0`	g

Bubble CO₂ for 3-4 min and adjust pH to be 6.5-6.8, if necessary. Then, add ferrihydrite suspension in the medium and dispense the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a CO₂ gas stream. Add appropriate amount of sulfur (powder). Seal the culture vessels with butyl rubber stoppers and autoclave at 105C for 1 hr.
Ferrihydrite:
Slowly titrate 320 ml of FeCl₃·6H₂O solution (60.0 g/L) with 10% (w/v) NaOH to pH 7.0 -7.5 with agitation on a magnetic stirrer (approximately 80-100 ml NaOH solution). Remove ferrihydrite into a container, cover with water and store at room temperature overnight. Remove supernatant after centrifugation at 2000 rpm for 5 min. Once again, wash ferrihydrite, centrifuge and remove supernatant. Finally, suspend ferrihydrite in 200 ml water.
Trace elements solution:

(NH₄)₂Fe(SO₄)₂·6H₂O	`78.4`	mg
CoCl₂·6H₂O	`23.8`	mg
NiCl₂·6H₂O	`16.6`	mg
Na₂MoO₄·2H₂O	`2.42`	mg
Na₂WO₄·2H₂O	`5.49`	mg
ZnSO₄·7H₂O	`14.4`	mg
CuCl₂·2H₂O	`0.17`	mg
Na₂SeO₄	`9.45`	mg
H₃BO₃	`0.62`	mg
MnCl₂·4H₂O	`19.8`	mg
SrSO₄	`0.92`	mg
CrK(SO₄)₂·12H₂O	`0.5`	mg
AlCl₃·6H₂O	`2.41`	mg
BaCl₂	`2.08`	mg
Na₂SiO₃·9H₂O	`14.2`	mg
KBr	`11.9`	mg
KI	`16.6`	mg
Na₂SO₄	`71.0`	mg
Distilled water	`1.0`	L